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. 2019 Sep 7;16(9):1304-1312.
doi: 10.7150/ijms.34617. eCollection 2019.

Mild hypothermia during the reperfusion phase protects mitochondrial bioenergetics against ischemia-reperfusion injury in an animal model of ex-vivo liver transplantation-an experimental study

Rui Miguel Martins  1 , João Soeiro Teodoro  2 , Emanuel Furtado  3 , Rui Caetano Oliveira  4 , José Guilherme Tralhão  5 , Anabela Pinto Rolo  2 , Carlos Marques Palmeira  2
Affiliations

Mild hypothermia during the reperfusion phase protects mitochondrial bioenergetics against ischemia-reperfusion injury in an animal model of ex-vivo liver transplantation-an experimental study

Rui Miguel Martins et al. Int J Med Sci. .

Abstract

The organ preservation paradigm has changed following the development of new ways to preserve organs. The use of machine perfusion to preserve organs appears to have several advantages compared with conventional static cold storage. For liver transplants, the temperature control provided by machine perfusion improves organ preservation. In this experimental study, we measured the effects of different temperatures on mitochondrial bioenergetics during the reperfusion phase. An experimental model of ex-vivo liver transplantation was developed in Wistar rats (Rattus norvegicus). After total hepatectomy, cold static preservation occurred at 4ºC and reperfusion was performed at 37ºC and 32ºC using a Langendorff system. We measured parameters associated with mitochondrial bioenergetics in the livers. Compared with the livers that underwent normothermic reperfusion, mild hypothermia during reperfusion caused significant increases in the mitochondrial membrane potential, the adenosine triphosphate content, and mitochondrial respiration, and a significant reduction in the lag phase (all P < 0.001). Mild hypothermia during reperfusion reduced the effect of ischemia-reperfusion injury on mitochondrial activity in liver tissue and promoted an increase in bioenergetic availability compared with normothermic reperfusion.

Keywords: adenosine triphosphate; bioenergetics; hypothermia; liver transplantation; mitochondria.

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Conflict of interest statement

Competing Interests: The authors have declared that no competing interest exists.

Figures

Figure 1
Figure 1
Schematic representation of reperfusion under hypothermic and normothermic conditions. Biopsies were taken at the end of the reperfusion time (A). The control group is not represented. Ten animals were analyzed per group.
Figure 2
Figure 2
Initial membrane potentials (Δψ) in the control group, group A (hypothermic reperfusion), and group B (normothermic reperfusion). The membrane potentials were determined in the presence of succinate as a respiratory substrate. Phosphorylation was induced by adding adenosine diphosphate (100 nmol). A statistically significant difference was found between groups A (hypothermic reperfusion) and B (normothermic reperfusion). **P < 0.01.
Figure 3
Figure 3
Lag phases in the control group, group A (hypothermic reperfusion), and group B (normothermic reperfusion) in the presence of succinate as a respiratory substrate. Phosphorylation was induced by adding adenosine diphosphate (100 nmol). A statistically significant difference was found between groups A (hypothermic reperfusion) and B (normothermic reperfusion). **P < 0.01.
Figure 4
Figure 4
The respiratory state 3 values for the control group, group A (hypothermal reperfusion), and group B (normothermic reperfusion). The respiratory status was determined in the presence of succinate. A statistically significant difference was found between groups A (hypothermic reperfusion) and B (normothermic reperfusion). **P < 0.01.
Figure 5
Figure 5
The respiratory control ratios in the control group, group A (hypothermal reperfusion), and group B (normothermic reperfusion). The respiratory control index was determined in the presence of succinate. A statistically significant difference was found between groups A (hypothermic reperfusion) and B (normothermic reperfusion). **P < 0.01.
Figure 6
Figure 6
Representative plot of the adenosine triphosphate (ATP) levels in the hepatic tissue of the control group, group A (hypothermic reperfusion), and group B (normothermic reperfusion). A statistically significant difference was found between groups A (hypothermic reperfusion) and B (normothermic reperfusion). **P < 0.01. ATP, adenosine triphosphate.
Figure 7
Figure 7
Hematoxylin and eosin (H&E)-stained sections of hepatic tissue from the hypothermic reperfusion group. The hepatic sinusoids do not show endothelial injury, and the hepatocytes contained normal intracellular organelles and nuclei, with no signs of apoptotic or necrosis (A: H&E 40×; B: H&E 400×).
Figure 8
Figure 8
Hematoxylin and eosin (H&E)-stained sections of hepatic tissue from the normothermic reperfusion group. The hepatic parenchyma architecture is preserved without lesions. There is moderate-to-severe disassociation of the hepatocytes. The hepatocytes contain normal nuclei and organelles, with no signs of necrosis or apoptosis (A: H&E 40×; B: H&E 400×).
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References

    1. Bertuzzo VR, Cescon M, Odaldi F, Di Laudo M, Cucchetti A, Ravaioli M. et al. Actual Risk of Using Very Aged Donors for Unselected Liver Transplant Candidates: A European Single-center Experience in the MELD Era. Annals of surgery. 2017;265:388–96. - PubMed
    1. Flores A, Asrani SK. The donor risk index: A decade of experience. Liver transplantation: official publication of the American Association for the Study of Liver Diseases and the International Liver Transplantation Society. 2017;23:1216–25. - PubMed
    1. Minambres E, Suberviola B, Dominguez-Gil B, Rodrigo E, Ruiz-San Millan JC, Rodriguez-San Juan JC. et al. Improving the Outcomes of Organs Obtained From Controlled Donation After Circulatory Death Donors Using Abdominal Normothermic Regional Perfusion. American journal of transplantation: official journal of the American Society of Transplantation and the American Society of Transplant Surgeons. 2017;17:2165–72. - PubMed
    1. Ruiz P, Gastaca M, Bustamante FJ, Ventoso A, Palomares I, Prieto M, Favorable Outcomes After Liver Transplantation With Normothermic Regional Perfusion From Donors After Circulatory Death: A Single-Center Experience. Transplantation; 2018. - PubMed
    1. Barbas AS, Goldaracena N, Dib MJ, Selzner M. Ex-vivo liver perfusion for organ preservation: Recent advances in the field. Transplantation reviews. 2016;30:154–60. - PubMed

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